Assessment of substrate inhibition of bacterial oligopeptidase B.

Malik Suliman Mohamed Mustafa, Yoshitaka Nakajima, Hiroshi Oyama, Nobuhisa Iwata, Kiyoshi Ito

Graduate School of Biomedical Sciences, Nagasaki University, 1–14 Bunkyo-machi, Nagasaki 852–8521, Japan.

Biological & pharmaceutical bulletin 2012

filter terms:

Oligopeptidase B (OPB; EC 3.4.21.83) from 2 Gram-negative bacteria, Stenotrophomonas maltophilia (Stm) and Serratia marcescens (Sem), and the Gram-positive bacterium Rhodococcus erythropolis (Re) were cloned and characterized to clarify their activities and substrate specificities using peptidyl-MCA substrates containing Arg or Lys. The cloned enzymes, Stm, Sem and ReOPBs, in addition to Escherichia coli OPB (EcOPB) were expressed using a pET expression system. Although the Stm and SemOPBs share 45% sequence identity to each other and up to 60% identity with respect to their catalytic domains, their activities towards MCA substrates were quite different. StmOPB is approximately 100-500 times more active than SemOPB and 3-30 times more active than EcOPB. The activity of ReOPB is comparable to that of StmOPB and it shares 40% and 36% identity to StmOPB and SemOPB, respectively. Some features of Stm, Re and EcOPBs are similar to those of previously cloned OPBs, which were also strongly inhibited by substrates, but SemOPB differs from all other OPBs in that it is not inhibited by substrates; even substrates containing double arginine at 35 µM did not inhibit SemOPB. On the other hand, the same substrates at only 5 µM inhibited the activity of the Stm, Re, and EcOPB. This phenomenon was not observed with substrates containing single or double lysine.

Citation

Malik Suliman Mohamed Mustafa, Yoshitaka Nakajima, Hiroshi Oyama, Nobuhisa Iwata, Kiyoshi Ito. Assessment of substrate inhibition of bacterial oligopeptidase B. Biological & pharmaceutical bulletin. 2012;35(11):2010-6